The invention relates first of all to a composition in the form of microcapsules or implants comprising a biodegradable polymeric or copolymeric excipient or a mixture of such excipients with an inherent viscosity of between 0.5 dl/g and 1.6 dl/g in CHCl3, and at least one active substance. The invention further relates to a composition in the form of microcapsules or implants comprising at least one biodegradable polymer or copolymer of high molecular weight and at least one water-soluble active substance of high specific surface area. Such compositions will be used to obtain a uniform release of the active substance over a prolonged period of up to more than three months.
These compositions, and especially the microcapsules, are mainly used in pharmacy, but can also be employed in other sectors, particularly in agrochemistry, i.e. in the plant protection sector.
The value of administering active principles in the form of sustained release compositions has been known for a long time, whether they be conventional pharmaceutical products, for example steroids, peptides or proteins (cf., for example, U.S. Pat. No. 3,773,919 to Boswell), or products for use in plant protection. The formulations adopted can take the form of microparticles in which the active principle is incorporated in a biodegradable polymer or copolymer such as a polylactide/co-glycolide copolymer (PLGA).
It has been found that, especially when a relatively constant or, in any case, uninterrupted release mode is soughtxe2x80x94this mode being referred to for example as xe2x80x9cmonophasexe2x80x9d in European Patent EP 58 481xe2x80x94PLGA-type polymers of relatively low molecular weight, i.e. of low viscosity, are required. European patents EP 21 234 (cf. Example 8.B.2. describing a copolymer of intrinsic viscosity 0.5 dl/g), EP 52 510, in which a copolymer with a viscosity of 0.38 dl/g in hexafluoro-isopropanol (HFIP) is tested in vivo, and EP 26 599, which describes, by way of example, polymers with viscosities of 0.12 to 0.20 dl/g and claims polymers with a viscosity of 0.08 to 0.30 dl/g, may be mentioned in this connection. The polymers described in said patents are presented as producing constant release compositions. The compositions of Patent EP 26 599 can contain fertility control agents, for example.
It is moreover important to note in this respect that in the opposition proceedings relating to European Patent EP 58 481, which are still in progress on the date of filing of the present patent application, the Applicant limited its main claim to polymers of low viscosity (below 0.3 or 0.5 dl/g), which, according to the Applicant, are the only ones capable of permitting a release of the monophase type.
Furthermore, when a longer release period, for example of more than one month, is sought, more complex problems appear and a solution proposed by Patent EP 0 302 582, for example, consists in mixing several types of microcapsules made up of polymers of different viscosities.
Now, the present Applicant has just found that certain polymers of high viscosity are suitable for the preparation of long-term sustained release compositions. It has also been found that the use of certain polymers produces compositions which have a very long-term monophase release profile without an initial period of no release (dead period). This applies particularly to polymers with an inherent viscosity preferably of at least 0.5 dl/g in CHCl3 and more preferably of at least 0.6 or 0.7 dl/g. In principle, however, the inherent viscosity of these polymers will not exceed 1.6 dl/g ,in CHCl3 and may be below 1.4 or 1.2 dl/g. Said polymers will preferably be PLGAs with a lactide/glycolide ratio varying from 40/60 to 90/10, and being preferably about 75/25.
The polymers according to the invention can be prepared by the customary methods, especially by opening of the lactide or glycolide rings. Such a process is described for example in U.S. Pat. No. 3,773,919.
In the present invention it is also possible to use a mixture of polymers of different high viscosities, but compositions containing only one polymer or copolymer are preferred.
The invention therefore relates first of all to a composition in the form of microcapsules or implants comprising a biodegradable polymeric or copolymeric excipient or a mixture of such excipients with an inherent viscosity of between 0.5 dl/g and 1.6 dl/g in CHCl3, and an active substance or a mixture of active substances, it being possible for these microcapsules or implants to release the active substance or mixture of active substances over a prolonged period of at least 1 month, preferably of at least 2 months and more preferably of at least 3 months.
Microcapsule is also understood to include microspheres, microparticles, nano-capsules, nanospheres or nanoparticles. Polymer will be understood as meaning a polymer, a copolymer or any mixture of these entities. Finally, active substance is understood as meaning an active substance, one of its salts, one of its precursors or any mixture of these compounds.
Salts of active substances which can be used for compositions according to the invention include especially the salts obtained from organic acids like acetic, malic, tartaric, oxalic, fumaric, citric, lactic, stearic, pamoic, methanesulphonic or p-toluenesulphonic acids, or from inorganic acids like hydrochloric, sulphuric, phosphoric or hydrobromic acids. It will be preferable to use a water-soluble product obtained by salification in the form of a cation, for example with acetic acid. However, it is possible to use an insoluble salt, for example a parnoate.
Particularly, the invention relates to a composition in the form of microcapsules or implants comprising a biodegradable polymeric or copolymeric excipient or a mixture of such excipients and an active substance or a mixture of active substances, said microcapsules or said implants being able to release the active substance or the mixture of active substances over a prolonged period of time of up to three months or more with an essentially monophase release profile, said composition being characterized in that:
either, when the composition is in the form of microcapsules:
either the viscosity of said polymers or copolymers is comprised between 0.7 dl/g and 1.6 dl/g in CHCl3 and the preparation process for said microcapsules does not comprise any stage of fusion of said microcapsules,
or the viscosity of said polymers ou copolymers is comprised between 0.5 dl/g and 1.6 dl/g in CHCl3 and said polymers or copolymers have an hydrophilic character;
or, when the composition is in the form of implants, the viscosity of said polymers or copolymers is comprised between 0.5 dl/g and 1.6 dl/g in CHCl3. Preferably, the viscosity of the polymers or copolymers for the compositions according to the invention will be at least equal to 0.9 dl/g in CHCl3.
The polymers or copolymers which can be used for the invention can be especially polymers such as those of lactic acid, glycolic acid, citric acid or malic acid, or else other biocompatible polymers like poly-xcex2-hydroxybutyric acid, polyorthoesters, polyorthocarbonates, poly-xcex1-cyanoacrylic acid esters, polyalkylene oxalates such as polytrimethylene or polytetramethylene oxalate, polyamino acids, etc. They can also be copolymers like PLGA, polystyrene, polymethacrylic acid, methacrylic acid/acrylic acid copolymers, polyamino acids, maleic anhydride polymers, ethyl cellulose, nitrocellulose, acetyl cellulose, etc. All these polymers or copolymers can be used by themselves or in any mixture. Generally, the PLGAs will comprise from 40 to 90% of lactide and from 10 to 60% of glycolide. It will be preferable to use D,L-PLGA and more preferable to use a D,L-PLGA produced from 70 to 80% of DL-lactide and 20 to 30% of glycolide. A PLGA synthesized from 75% of DL-lactide and 25% of glycolide will be particularly suitable for the invention.
Another particularly preferred polymer for the invention is L-PLGA obtained from L-lactide and glycolide. Compared with D,L-PLGA of the same viscosity, L-PLGA assures a slower release and represents an alternative to the D,L-PLGAs of higher viscosity.
In a general manner, the polymers or copolymers possessing a hydrophilic character will be preferred. Hence, preference will generally be given to the PLGAs obtained by ring opening with hydrophobic initiators such as those of the lauryl alcohol type, those obtained by ring opening with hydrophilic initiators such as those of the lactic acid or glycolic acid type.
By polymer or copolymer possessing a hydrophilic character, it is meant a polymer or copolymer for which the terminal chain is polar (for example, this terminal chain includes at its end an acidic function), by opposition to a polymer or copolymer possessing a hydrophobic character for which the terminal chain is apolar (for example, this terminal chain is an aliphatic chain).
The acid number, which corresponds to the number of milliequivalents of KOH required per gram of polymer to neutralize the free acidity, seems to be the parameter which correlates best with the hydrophilic or hydrophobic character of a polymer or copolymer. Wherever the terminal chains of the polymers or copolymers may include a free acidic function, owing to the nature of the monomer, this acid number may be measured.
In a general manner, the Applicant has found that the hydrophilic polymers produce a better release profile. Hence, the acid number of the polymers used for the invention will preferably be at least equal to 1, or better, 1.2, and more preferably at least equal to 1.5 or 2.
The core loading of the microcapsules according to the invention, i.e. the ratio of the weight of encapsulated pure peptide to the total weight of the microcapsule, will generally be between 0 and 20% and preferably between 2 and 15%. In the case of triptorelin acetate, the loading will preferably be less than or equal to 10% and more preferably between 4 and 8% for forms which permit release over a period of about 3 months. In the case of lanreotide acetate, the loading will preferably be between 10 and 20%.
In the case of implants, the core loading will generally be between 0 and 30% and preferably between 15 and 25%.
The encapsulation step can be a so-called coacervation step, such as that described in U.S. Pat. No. 3,773,919 or European Patent EP 52 510.
It is also possible to use a so-called melting-extrusion process such as that described in European Patent EP 58 481 or U.S. Pat. No. 5,225,205, the products obtained then optionally being ground by the customary methods to give microparticles.
In another respect, a water-soluble active principle such as a water-soluble salt of a peptide, for example the acetate, can be used. It is also possible to use an insoluble salt of a soluble molecule, such as a fatty acid salt of a peptide, for example a peptide pamoate such as that described in British Patent GB 2 209 937.
The compositions obtained by melting-extrusion using the polymers according to the invention can also be presented in the form of implants and be used as such.
These implants are preferably small (mini-implants or microimplants) with a diameter of the order of 1 mm, for example of between 0.8 and 1.2 mm. The length of these implants can be for example between 10 and 35 mm, for example of the order of 25 mm. These implants give very advantageous results with low doses of active principle, for example of the order of 3 mg of triptorelin acetate per implant. Such implants can release the active principle over a period of up to 3 months.
Moreover, it has been found that the form of the active principle can also influence the diffusion of this product. In particular, if an active principle can be obtained in a crystalline or amorphous form, it is not arbitrary which of the two forms is chosen.
Patent application EP 709 085 describes microcapsules comprising a polymer and an amorphous water-soluble active substance. It is particularly concerned with the importance of obtaining small particles of active substance, preferably with a size of less than 10 m. However, this patent application does not disclose any process for the preparation of said particles and no mention is made of the effect of the specific surface area of the particles of active principle on the release profile of the compositions containing these particles. Now, the Applicant has already been using, since 1986, microcapsules containing an amorphous active substance, namely triptorelin acetate, sold under the name Decapeptyl 3.75 mg, which has a particle size of only about 8 m. However, it has found that the particle size is not the only determining parameter for favouring release over a prolonged period of up to more than three months or more.
In principle, the question of amorphous character does not arise for products such as peptides or proteins, whose method of preparation, especially lyophilization, leads to an amorphous product in the majority of cases, as for Decapeptyl 3.75 mg.
There are copious illustrations of this phenomenon in the literature and the following articles may be mentioned in particular: Hsu, C.C. et al., Pharmaceutical Research, 12 (1), 69-77 (1995) or Towns, J. K., Journal of Chromatography, A, 705 (1), 115-27 (1995).
The invention therefore also relates to a composition in the form of microcapsules or implants comprising at least one biodegradable polymer or copolymer of high molecular weight and at least one water-soluble active substance of high specific surface area. Particularly, said specific surface area is greater than 2 m2/g, and preferably greater than 3 m2/g. More preferably, said specific surface area is greater than 5 m2/g or 10 m2/g. Even more preferably, said specific surface area is greater than 20 m2/g, and preferably greater than 30 m2/g.
The invention relates preferably to the above compositions in which the water-soluble active substance is a protein or a peptide.
It further relates to the above compositions for which the polymer or copolymer viscosity is comprised between 0.5 and 1.6 dl/g in CHCl3, and preferably comprised between 0.9 and 1.6 dl/g in CHCl3. Particularly, polymers or copolymers of viscosity comprised between 0.7 and 1.3 dl/g in CHCl3 may be chosen and more preferably polymers or copolymers of viscosity comprised between 0.7 and 1.3 dl/g. PLGAs are particularly adapted for the invention. Preferably, said PLGAs will be produced from 40 to 90% lactide and 10 to 60% glycolide, and more preferably from 70 to 80% lactide and 20 to 30% glycolide. The hydrosoluble active substances incorporated in the microcapsules or implants will preferably be proteins or peptides.
Compositions comprising an active substance of high specific surface area will preferably be such that viscosity of the polymer or copolymer is comprised between 0.5 and 1.6 dl/g in CHCl3 and that the polymer or copolymer presents an hydrophilic character, the acid number of the latter being greater than 1 meq KOH per gram of polymer or copolymer, and preferably greater than 1.2, more preferably 1.5 mEq or even 2 meq KOH per gram of polymer or copolymer.
The invention further relates to compositions in the form of microcapsules or implants comprising an active substance of high specific surface area characterized in that the polymer or copolymer is a PLGA, and preferably a PLGA produced from 70 to 80% lactide and 20 to 30% glycolide, the viscosity of said PLGA being comprised between 0.5 and 1.6 dl/g on CHCl3 and the active substance incorporated in the microparticles or implants being a protein or a peptide.
These microcapsules or implants permit a monophase release profile in which the initial peak (or burst) is reduced in comparison with certain other preparations using a polymer of lower molecular weight, so they make it possible to release the active substance over a prolonged period of up than three months or more.
In other words, the Applicant has found that the release properties, notably the monophase type release, of compositions in the form of microcapsules or implants, particularly of compositions based on PLGA and including as an active principle a peptide or a protein, are considerably improved if at least one of the following characteristics is present:
a) the polymer or copolymer is a PLGA which presents a viscosity in chloroform of at least 0.5 dl/g, preferably of at least 0.9 dl/g and in principle lower than 1.6 dl/g;
b) the polymer or copolymer is a PLGA which is prepared from 70 to 80% lactide and from 20 to 30% glycolide;
c) the polymer or copolymer presents an hydrophilic character, and preferably has an acid number greater than I mEq KOH, and m ore preferably greater than 1.2 or even 1.5 mEq KOH per gram of polymer or copolymer;
d) the active principle, preferably a peptide or a protein, has a high specific surface area and greater than 2 m2/g, preferably greater than 10 m2/g, more preferably greater than 20 m2/g or even greater than 30 m2/g;
these characteristics being optionally combined with the use of a L-PLGA instead of a D,L-PLGA.
According to its present knowledge, the Applicant is of the opinion that characteristic d) taken alone is very important-~and may be advantageously combined to the other characteristics a) b) or c). Particularly, the characteristic d) may be combined to following characteristics : a) alone, b) alone, c) alone, a) and b) together, a) and c) together, b) and c) together, or a), b) and c) together. More preferably, characteristic d) will be combined at least with characteristic c).
Among the active substances which can be used for the different aspects of the invention, there may be mentioned in particular proteins and peptides. Said active substances can be selected for example from the group consisting of the following substances: triptorelin or one of its salts, particularly triptorelin acetate, lanreotide or one of its salts, particularly lanreotide acetate, octreotide or one of its salts (as described for example in European Patent EP 29 579), particularly octreotide acetate or pamoate, a compound with LH-RH activity, such as triptorelin, goserelin, leuprorelin, buserelin or their salts, an LH-RH antagonist, a GPIIb/IIIa antagonist, a compound with a similar activity to a GPIIb/IIIa antagonist, erythropoietin (EPO) or one of its analogues, the various types of interferon-xcex1, interferon-xcex2, or -xcex3, somatostatin, a somatostatin derivative such as that described in European Patent EP 215 171, a somatostatin analogue such as that described in U.S. Pat. No. 5,552,520 (this patent itself includes a list of other patents describing somatostatin analogues, which are incorporated in the present patent application by way of reference), insulin, a growth hormone, a growth hormone releasing factor (GRF), a growth hormone releasing peptide (GHRP), an epidermal growth factor (EGF), a melanocyte stimulating hormone (MSH), a thyrotropin releasing hormone (TRH) or one of its salts or derivatives, a thyroid stimulating hormone (TSH), a luteinizing hormone (LH), a follicle stimulating hormone (FSH), a parathyroid hormone (PTH) or one of its derivatives, a lysozyme hydro chloride, a parathyroid hormone related peptide (PTHrp), an N-terminal peptide fragment (position 1xe2x86x9234) of human PTH hormone, vasopressin or one of its derivatives, oxytocin, calcitonin, a calcitonin derivative with a similar activity to that of calcitonin, a calcitonin gene related peptide (CGRP), glucagon, a peptide similar to glucagon (GLP), gastrin, a gastrin releasing peptide (GRP), secretin, pancreozymin, cholecystokinin, angiotensin, human placental lactogen, human chorionic gonadotropin (HCG), enkephalin, an enkephalin derivative, colony stimulating factor (CSF), endorphin, kyotorphin, interleukins, for example interleukin-2, tuftsin, thymopoietin, thymosthymlin, thymic humoral factor (THF), thymic serum factor (TSF), a derivative of thymic serum factor (TSF), thymosin, thymic factor X, tumour necrosis factor (TNF), motilin, bombesin or one of its derivatives as described in U.S. Pat. No. 5,552,520 (this patent itself includes a list of other patents describing bombesin derivatives, which are incorporated in the present patent application by way of reference), prolactin, neurotensin, dynorphin, caerulein, substance P, urokinase, asparaginase, bradykinin, kallikrein, nerve growth factor, a blood clotting factor, polymixin B, colistin, gramicidin, bacitracin, a protein synthesis stimulating peptide, an endothelin antagonist or one of its salts or derivatives, a vasoactive intestinal polypeptide (VIP), adrenocorticotropic hormone (ACTH) or one of its fragments, a platelet derived growth factor (PDGF), a bone morphogenetic protein (BMP), a pituitary adenylate cyclase activating polypeptide (PACAP), neuropeptide Y (NPY), peptide YY (PYY), a gastric inhibitory polypeptide (GIP) and polynucleotides, especially double-stranded RNAs (ds-RNAs) such as those described in Patent application EP 0 300 680 or French Patent no. 2 622 586.
ds-RNA is preferably understood as meaning polyadenylic acid complexed with polyuridylic acid, which is also called poly(A)-poly(U) or Poly-adenur. Other ds-RNAs can be used for the invention, especially a complex of polyinosinic acid with polycytidylic acid, which is also known by the name poly(I)-poly(C), as well as these same complexes modified by the introduction of uridylic acid into the polycytidylic acid chain, such as the product Ampligen from the HEMISPHERx company (for a description of these products, reference may be made especially to European Patent application EP 0 300 680). The ds-RNA used can be for example a mixture of ds-RNAs as defined above. The ds-RNAs are preferably prepared by the process described in French Patent no. 2 622 586.
A high specific surface area can be obtained for the previously mentioned substances as soon as they are water-soluble or transformed into water-soluble substances, for example by salification or grafting of a water-soluble chain on their structure. This is particularly valid for the previously mentioned peptides and proteins. Any other water-soluble active substance or one of its salts or precursors, and particularly the salts obtained by salification with acetic acid, may also be used by a person skilled in the art for this aspect of the invention if they consider it appropriate.
According to one of the preferred aspects of the invention, the peptide or protein with a high specific surface area are chosen from the group consisting of triptorelin acetate, lanreotide acetate or octreotide acetate.
Peptide and/or protein are understood in the present application as meaning both the peptide and/or the protein themselves and pharmacologically active fragments, salts or derivatives of these peptides or proteins.
The water-soluble active substance as used to manufacture microcapsules or implants according to the invention, and particularly triptorelin acetate, lanreotide acetate, octreotide acetate, goserelin, leuprorelin, buserelin or their salts, is preferably obtained by a process which principally involves two steps:
a lyophilization step comprising the rapid immersion of a dilute solution of the water-soluble substance in a medium whose temperature is below xe2x88x9250xc2x0 C., and preferably below xe2x88x9270xc2x0 C.; and
optionally a grinding step, which will preferably comprise ultrasonic grinding.
Dilute solution of the active substance is understood as meaning a solution whose concentration of said active substance is less than half the saturation concentration and preferably less than a quarter of said saturation concentration when the latter is at least equal to at least 200 g/l. This process produces an active substance of high specific surface area.
Rapid immersion must be understood as meaning contact with a low temperature medium, causing instantaneous freezing of the solution of water-soluble substance.
For the lyophilization, the solution may be frozen for example in a tray floating in a tank of liquid nitrogen, before the actual lyophilization is carried out.
Preferably, in order to obtain a maximum specific surface area, the rapid immersion of the solution will be preceded by a micronization of the solution of active substance. When the solution of active substance is micronized beforehand, the temperature of the low temperature medium may only be below xe2x88x9250xc2x0 C.
For example, to obtain a very high specific surface area, it may be chosen to atomize the solution by spraying it through an atomizer onto a metal plate at very low temperature. The temperature of the plate will preferably be below xe2x88x9250xc2x0 C. and more preferably below xe2x88x9270xc2x0 C. or even xe2x88x9280xc2x0 C. or xe2x88x92120xc2x0 C. This temperature may be reached for example by immersing a metal plate in a very low temperature medium, for example liquid nitrogen. According to one preferred variant of the invention, the metal plate is hollow and the solution is sprayed inside said plate by means of an atomizer.
Other freezing techniques can be considered, for example-atomization of the solution of active substance into a precooled bath of a non-solvent for said active substance. The non-solvent will preferably be a liquefied gas, such as for example liquid nitrogen.
Another possibility is to freeze the active substance solution on a rotating plate (drum freezing). As previously indicated, this freezing will preferably be preceded by a micronization of the active substance solution.
When the process of freezing in a tray is applied to an active substance in order to prepare sustained release microcapsules or implants according to the invention, the specific surface area of the active substance, after lyophilization but before grinding, will preferably be greater than 2 m2/g. The specific surface area of the active substance will more preferably be greater than 3 m2/g or even 5 m2/g.
If a specific surface area greater than 10 m2/g is required, the process which includes a micronization step will preferably be employed. The specific surface area obtained for the active substance after lyophilization will preferably be greater than 15 m2/g. This specific surface area will even more preferably be greater than 20 m2/g or even 30 m2/g.
The specific surface areas obtained may be varied by varying the freezing conditions of the solution of active substance by way of different parameters, such as for example the freezing rate or the concentration of the solution.
The specific surface area of the active substance is a favourable factor for obtaining release over a prolonged period, particularly in the case of microcapsules. In fact, as already mentioned, particles of an active substance which have the same size but different specific surface areas will give totally different results with the same polymeric excipient.
The invention therefore also relates to the processes as described above, applied to a biologically active water-soluble substance. It further relates to the biologically active water-soluble substance as obtained by these processes, said substance having a high specific surface area.
Particularly, the invention relates to triptorelin acetate, lanreotide acetate or octreotide acetate as obtained by the previously described processes, or to a double strand RNA, preferably polyadenylic acid complexed with polyuridylic acid as obtained by these processes.
As indicated above, the compositions according to the invention are preferably used in the pharmaceutical sector. The pharmaceutical compositions can be administered to a patient by different routes, however, the preferred route is subcutaneous or intra-muscular injection. The microcapsules according to the invention can first be suspended in a vehicle appropriate for injection, such as an aqueous solution of sodium chloride or an aqueous solution of mannitol.
Unless defined otherwise, all the technical and scientific terms used here have the same meanings as those commonly understood by an ordinary specialist in the field to which this invention belongs. Likewise, all the publications, patent applications, patents and any other references mentioned here are incorporated by way of reference.